Journal: Cell Death Discovery
Article Title: Bridging the marrow: a co-culture-platform of leukemia cells and MS5-derived stromal cells or adipocytes
doi: 10.1038/s41420-025-02631-5
Figure Lengend Snippet: A and B B-precursor ALL cell lines EU3, KOPN-8, RCH-ACV, Nalm6, Reh, and RS4;11 (each dot represents one cell line) were cultured alone or together with stromal cells or adipocytes derived from MS5 cells. Cell viability ( A ) and the number of viable cells per well ( B ) were determined after 7 days by FACS. C EU3 mono-culture was compared to co-culture with MS5-derived stromal cells or adipocytes cultured in DMEM/F12 medium without FCS, compared to medium with increasing concentrations of FCS as indicated. Viability and the number of viable cells per well were determined by FACS analysis after 7 days. D The B-precursor ALL cell line RS4;11 was labeled with CellTrace Violet (CT) and cultured alone or together with MS5-derived stromal cells or adipocytes, either directly or in transwell inserts. Viability and mean fluorescence intensity (MFI) of CellTrace Violet were determined on day 7 after seeding. E RS4;11 cells were co-cultured in transwell inserts over medium (mono-culture), MS5-derived stromal cells, or adipocytes for 72 h. Subsequently, ATP production was analyzed by the Seahorse Extracellular Flux Analyzer. F MS5-derived stromal cells (left panel) and adipocytes (right panel) were incubated with leukemia-conditioned media for 24 h and subsequently analyzed by the Seahorse Extracellular Flux Analyzer. ATP production rates are depicted. Data are presented as mean ± SEM ( A , B , D ) or as mean ± SD ( C , E , F ) of at least three independent experiments performed in triplicate ( A – C ) or duplicates ( D ) or in three to five technical replicates ( E , F ). RM one-way ANOVA with Tukey’s multiple comparisons test ( A , B ), two-way ANOVA with Dunnett’s ( C , E , F ) or Tukey’s ( D ) multiple comparisons test, * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.
Article Snippet: B-precursor ALL cell lines Nalm6 (DSMZ no. ACC 128; Species: human), Reh (DSMZ no. ACC 22; Species: human), RS4;11 (DSMZ no. ACC 508; Species: human), RCH-ACV (DSMZ no. ACC 548; Species: human), EU3 (also known as cell line 697; DSMZ no. ACC 42; Species: human), and KOPN-8 (DSMZ no. ACC 552; Species: human) were maintained in RPMI (Gibco, Thermo Fisher Scientific) supplemented with 10–20% FCS and 2% L -glutamine.
Techniques: Cell Culture, Derivative Assay, Co-Culture Assay, Labeling, Fluorescence, Incubation